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哈佛医学院的转基因RNA干扰项目:资源与验证

The Transgenic RNAi Project at Harvard Medical School: Resources and Validation.

作者信息

Perkins Lizabeth A, Holderbaum Laura, Tao Rong, Hu Yanhui, Sopko Richelle, McCall Kim, Yang-Zhou Donghui, Flockhart Ian, Binari Richard, Shim Hye-Seok, Miller Audrey, Housden Amy, Foos Marianna, Randkelv Sakara, Kelley Colleen, Namgyal Pema, Villalta Christians, Liu Lu-Ping, Jiang Xia, Huan-Huan Qiao, Wang Xia, Fujiyama Asao, Toyoda Atsushi, Ayers Kathleen, Blum Allison, Czech Benjamin, Neumuller Ralph, Yan Dong, Cavallaro Amanda, Hibbard Karen, Hall Don, Cooley Lynn, Hannon Gregory J, Lehmann Ruth, Parks Annette, Mohr Stephanie E, Ueda Ryu, Kondo Shu, Ni Jian-Quan, Perrimon Norbert

机构信息

Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115.

Boston University, Boston, Massachusetts 02215.

出版信息

Genetics. 2015 Nov;201(3):843-52. doi: 10.1534/genetics.115.180208. Epub 2015 Aug 28.

Abstract

To facilitate large-scale functional studies in Drosophila, the Drosophila Transgenic RNAi Project (TRiP) at Harvard Medical School (HMS) was established along with several goals: developing efficient vectors for RNAi that work in all tissues, generating a genome-scale collection of RNAi stocks with input from the community, distributing the lines as they are generated through existing stock centers, validating as many lines as possible using RT-qPCR and phenotypic analyses, and developing tools and web resources for identifying RNAi lines and retrieving existing information on their quality. With these goals in mind, here we describe in detail the various tools we developed and the status of the collection, which is currently composed of 11,491 lines and covering 71% of Drosophila genes. Data on the characterization of the lines either by RT-qPCR or phenotype is available on a dedicated website, the RNAi Stock Validation and Phenotypes Project (RSVP, http://www.flyrnai.org/RSVP.html), and stocks are available from three stock centers, the Bloomington Drosophila Stock Center (United States), National Institute of Genetics (Japan), and TsingHua Fly Center (China).

摘要

为推动果蝇的大规模功能研究,哈佛医学院(HMS)设立了果蝇转基因RNA干扰项目(TRiP),并制定了几个目标:开发能在所有组织中起作用的高效RNA干扰载体;在社区的参与下,构建一个基因组规模的RNA干扰品系库;通过现有的品系中心分发已构建好的品系;使用逆转录定量PCR(RT-qPCR)和表型分析尽可能多地验证品系;开发用于识别RNA干扰品系并检索其质量现有信息的工具和网络资源。鉴于这些目标,我们在此详细描述我们开发的各种工具以及该品系库的现状,目前该品系库由11,491个品系组成,覆盖了71%的果蝇基因。关于这些品系通过RT-qPCR或表型进行特征描述的数据可在一个专门的网站——RNA干扰品系验证与表型项目(RSVP,http://www.flyrnai.org/RSVP.html)上获取,品系可从三个品系中心获得,即美国的布鲁明顿果蝇品系中心、日本的国立遗传学研究所和中国的清华果蝇中心。

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